High frequency regeneration from leaf derived callus and its histological studies in Lepianthes umbellata

Abstract

ABSTRACTA protocol was standardized for callus initiation and shoots regeneration from leaf explants of Lepianthes umbellata. The explant was cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 2,4-Dichlorophenoxyacetic acid (2,4-D), 6-Benzylaminopurine (BAP), and 1-Naphthalene acetic acid (NAA) for shoot bud initiation and multiplication. High frequency of shoot regeneration and maximum number of shoots per explant were achieved on MS medium fortified with 0.5 mg L−1 NAA and 1.0 mg L−1 BAP. The elongations of shoots were achieved on the same medium supplemented with 1.0 mg L−1 Gibberelic acid (GA3), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA) at different concentrations on half strength MS medium were used for rooting of the micro shorts. Highest efficiency for rooting was observed on MS with 1.5 mg L−1 IBA. Regenerated plantlets were successfully transferred for acclimatization (70%). Histological studies showed that embryoids and shoot buds initiating from the callus masses, that is, indirect organogenesis. Cytological variations were observed in callus cultures.